棉花SUPERMAN类锌指蛋白基因GZFP的启动子及功能分析  

Functional Analysis of the SUPERMAN-like Zinc Finger Protein Gene GZFP and Its Promoter

作　　者：杨郁文[1] 周建武[2] 张保龙[1] 范晓慧[1] 任永哲[1] 陈天子[1] 

机构地区：[1]江苏省农业科学院农业生物技术研究所/江苏省农业生物学重点实验室,江苏南京210014 [2]扬州大学生物科学与技术学院,江苏扬州225009

出　　处：《棉花学报》2011年第6期483-489,共7页Cotton Science

基　　金：转基因生物新品种培育重大专项(2009ZX08005-003B);江苏省自主创新项目(2009ZX08005-003B)

摘　　要：GZFP是来源于陆地棉的一个SUPERMAN类锌指蛋白基因。为了进一步了解该基因功能,本研究通过染色体步移(Genome walking)获得其启动子区域,并构建该片段与GUS连接的重组载体pBI-pGZFP-5::GUS,通过花浸染法转化拟南芥。转基因植株的GUS染色结果表明GUS基因主要在根部以及花器官表达,而叶片中的表达量很低,并且根部的表达量在整个生育期都很强。通过半定量RT-PCR分析不同诱导下GZFP基因的表达变化,发现ABA、干旱、盐胁迫诱导可以提高其表达量。构建GZFP过量表达载体并通过叶盘法转化烟草,烟草转化株的表型无明显变化,但是转化株中NtOPBP1和NtERD10的表达量较未转化株明显增强,而这两个基因都参与了植物的抗逆反应,说明GZFP基因可能与植物抗逆相关。GZFP, derived from Gossypium hirsutum L, belongs to the SUPERMAN-like zinc finger protein. To further analyze the function of GZFP, the promoter region of GZFPwas obtained by genome walking. The recombination vector pBI-pGZFP-5：：GUS containing the promoter was constructed and transformed to Arabidopsis by Agrobacterium-mediated method. GUS staining showed GZFP promoter driven GUS activity was specifically detected in roots and flowers and the activity was very low in leaves. In addition, the GUS activity in roots was very strong throughout the whole growth period. By semi-quantitative RT-PCR, we have found the expression of GZFP was induced by ABA, drought and salt. The over expression vector was con- structed by inserting GZFP into the pCAMBIA2301, tobacco plants were transformed by co-cultivating leaves method via Agrobacterium mediation. The object gene was verified to have been integrated into the genome of tobacco by PCR. The phenotype of transgenic plants was normal compared to the wild type, but the expression of NtOPBP1 and NtERDIO was relatively higher than the wild type. As the two genes involved in the stress reaction, GZFP may have some relevance to the plant stress reaction.

关 键 词：锌指蛋白 陆地棉 启动子 抗逆 

分 类 号：S562[农业科学—作物学]